[Carrier frequency of alpha thalassemia mutations among newborns in northern Iran]

Background and Objective: Alpha Thalassemia is one of the most prevalent hemaglobinophaties worldwide. Alpha thalasseima patients may represent wide spectrum of symptoms ranging from asymptomatic to severe life threatening anemia. This study was done to assess the carrier frequency of alpha globin gene mutations among newborns in north of Iran

Methods: In this descriptive study, 412 cord blood samples of neonate from Amir Mazandari hospitali were randomly selected during 2012. Genomic DNA was extracted using phenol-chloroform method. Multiplex Gap- PCR and PCR-RFLP methods were applied in order to detect three common gene deletions, one triplication and one point mutation

Results: Total allelic frequency of investigated mutations was 0.0825. The -alpha3.7 deletion with allelic frequency of 0.0485 was the most prevalent mutation among 412 neonates. Allelic frequencies of -alpha4.2, alphaalphaalphaanti3.7 triplication and alpha-5nt mutations were 0.0206, 0.0109 and 0.0024; respectively and -Med double gene deletion was not detected

Conclusion: Most mutated cases had single gene deletion that is asymptomatic while -Med double gene deletion was not detected among the neonates. Therefore, there is low probability of a child birth with Hb H disorder in the region

[Detection of common mutations causing drug resistance to mycobacterium tuberculosis strains among patients with tuberculosis using line probe assay method]

Drug resistance to tuberculosis and especially multiple drug resistance tuberculosis [MDR-TB] variants are a serious problem in tuberculosis patients and make difficulties in controlling the disease. This study was coducted for detection of common mutations causing drug resistance of mycobacterium tuberculosis strains among tuberculosis patients using line probe assay method. In this descriptive study, fifty four sputum samples of tuberculosis patients were randomly selected in health centers of Mazandaran, northern Iran during 2012. After culturing of sputum samples on Lowenstein-Jensen medium, genomic DNA was extracted from colonies using CTAB method. Molecular analysis of mutations causing resistance to five different antibiotics including Isiniazide, Rifampin, Sterptomycine, Amicasin / Canamycine, Kinolon were performed using long probe assay [LPA] method. Out of 54 sputum samples, three [5.5%], three [5.5%], four [7.4%] were resistance to Kinolon, Amicasin / Canamycine and Sterptomycine, respectively. Mutation in KATG was seen in 2 samples resistant to Isiniazide. Mutation in rpoB 516 was seen in 3 samples resistant to Rifampin. Four samples [7.4%] were resistant to the two anti-tuberculosis antibiotics, while three samples were resistant to Sterptomycine and Kinolon and one sample was resistant to Rifampin and Canamycine. 7.4% of sputum samples were resistant to the two anti- tuberculosis antibiotics. Line probe assay is a rapid and suitable method for detecting tuberculosis drug resistance

Beta globin gene haplotypes associated with hemoglobin D-Punjab in northern Iran

Background and Objective: Hemoglobin D-Punjab is one of the variant of hemoglobin caused by a mutation on position 121 of beta globin gene which is frequent in India, PakistanandIran. Heterozygote form of this variant is mainly asymptomatic while in combination with hemoglobin S, severe form of anemia occure. This study was carried out to determine the beta globin gene haplotypes associated with hemoglobin D-Punjab in Northern Iran. Methods: This descriptive study was carried out on families of 18 individuals whom were carriers of hemoglobin D-Punjab in Sari in Northern Iran. Genomic DNA was extracted from peripheral blood samples using Phenol-chloroform st and ard protocol. In order to identify different haplotypes associated with hemoglobin D-Punjab, PCR-RFLP method and family linkage analysis were used. Results: In 17 subjects hemoglobin D-Punjab was linked to [+ - - - - + +] haplotype and in one case association with [- + + - + + +] haplotype was observed. Conclusion: The hemoglobin D-Punjab alleles have mainly unicentric origin and [- + + - + + +] rare haplotype may have different genetic origin or is created as a result of gene recombination

[Comparison of fluorescent spot test, decolorization test and quantitative enzyme assay in detection of G6PD enzyme deficiency]

Background and Objective: The national screening program for G6PD enzyme deficiency is not able to detect all affected neonates. This study was done to compare the fluorescent spot test [FST], decolorization test, and quantitative enzyme assay [QEA] for detecting G6PD enzyme deficiency in neonates

Methods: In this descriptive study, cord blood samples of 365 neonates were collected. Decolorization test, QEA and DNA test was done for each sample. All of the neonates were tested by FST as a part of national screening program on heel-prick blood sample collected on day 3-5 after birth. QEA was considered as the gold standard. According to QEA test results, neonates with <20% and 20-60% of mean normal enzyme activity were considered as total deficient and partial deficient, respectively

Results: Fluorescent spot test detected 13 male neonates with G6PD enzyme deficiency while decolorization test identified 18 male and 1 female neonates. Using QEA, 19 of male and 28 of female neonates with G6PD enzyme deficiency [26 cases with partial and 2 cases with total deficiency] were diagnosed. DNA analysis detected 34 female case as heterozygote and 14 male neonates as hemizygote for the disease

Conclusion: Fluorescent spot test do not have required sensitivity for screening of neonates with G6PD enzyme deficiency. QEA test is recommended to replace the fluorescent spot test in national screening program

[Prevalence of alloimmunization in major beta thalassemia in northern Iran [2010]]

Repeated blood transfusion is the major treatment for patients with major thalassemia. However due to antigen encounters, it may initiate body reactions, including alloantibodies against red blood cell antigens. This study was done to determine the Prevalence of alloimmunization in major beta thalassemia patients in northern Iran. This descriptive - analytic study was carried out on 218 thalassemic patients [100 males and 118 females] with average age of 22.5 +/- 7 years in northern Iran during 2010. Each sample was tested for the presence of Alloantibodies including C, Cw, Lea, E, Lua, Leb, K, Jkb, N, P1, D, Jka, M, S, Xga, e, Fya, s, c, Fyb, k, Kpa, Jsb, Lub and Coa. Eighty eight cases [40.4%; 95% CI: 33.9-46.9] were positive for the presence of alloantibodies. Alloantibodies against C, Cw, Lea red blood cell surface antigens were the most prevalent [40%]. No significant correlation was found between emergence of alloantibody with the age of initial, frequency and duration of blood transfusion. Alloimmunization is a common observation in thalassemic patients and should be prevented by transfusing compatible blood

Diagnostic value of fructosamine and glycosylated hemoglobin in estimating blood glucose level in diabetic patients with thalassemia major

Diabetes mellitus is one of complications that thalassemia major patients face with. Hence, blood glucose monitoring is of vital importance to these patients. Because of high level of fetal hemoglobin in these patients, the measurement of hemoglobin AI[c] is not reliable and should be displaced by fructosamine test. The current descriptive study was carried out on 33 beta-thalassemia major patients afflicted with diabetes mellitus [21 female and 12 male cases]. Blood glucose level, fructosamine, hemoglobin AI[C], serum ferritin and fetal hemoglobin were measured. Blood glucose levels are 204 +/- 103 mg/dL and 221 +/- 101 mg/dL [p=0.63]; fetal hemoglobin levels are 9% +/- 7% and 13% +/- 9% [p=0.22]; serum ferritin levels are 1744 +/- 1534 ng/mL and 3253 +/- 1773 ng/mL [p=0.96] in female and male patients, respectively. The level of fructosamine [42 +/- 124 mmol/L] and glycosylated hemoglobin [8.9% +/- 1.8%] are correlated significantly [r=0.69, p<0.01]. Both Hemoglobin A[1c] [r=0.75, p<0.01] and fructosamine [r=0.54, p<0/01] show a significant correlation with blood glucose level. In diabetic thalassemia major patients with frequent blood transfusion, the level of fructosamine and glycosylated hemoglobin are related significantly, therefore; they can be used alternatively

[Molecular evaluation of hemoglobin D mutations in Mazandaran province, Iran]

Hemoglobinopathies are among the most prevalent genetic disorders worldwide, and occur as a result of mutations in the gene involved in synthesizing hemoglobin chains. By now more than 1000 defects in hemoglobin chains are discovered. Hemoglobin D [Hb D] is one of these disorders, identified by a single nucleotide mutation on codon 121 of beta globin chain. This study was carried out to evaluate Hb D mutations through molecular methods in Mazandaran province of Iran. This descriptive laboratory study was done on 70 patients with an electrophoresis band in hemoglobin-S zone in Mazandaran province of Iran during 2010-11. Capillary zone electrophoresis was done to find out Hb D in 51 patients. Subsequently, PCR-RFLP was performed to evaluate the samples at molecular level. Molecular investigation revealed all cases are carriers of hemoglobin D-Punjab. Two patients were shown to be homozygote carriers of the abnormal gene. This study showed all Hb D affected patients were carriers of Hb D Punjab

Short-term exposure to high levels of natural external gamma radiation does not induce survival adaptive response

In some areas of Ramsar, a city in northern Iran, residents receive a much higher annual radiation exposure than is permitted for radiation workers. Induction of adaptive response in residents of Ramsar has been reported previously. In this study induction of such a response in short term exposure to high background levels of gamma radiation is investigated. Fifty male NMRI mice were randomly divided into four groups of 10-17 animals and 53 Wistar rats were randomly divided into five groups of 10-12 animals were studied. Animals in the 1[st] group were kept for 7 days in an outdoor area with normal background radiation while the 2[nd], 3[rd], 4[th] and 5[th] [in case of rats] groups were kept in 3 different outdoor areas with naturally elevated levels of gamma radiation. Animals were then exposed to a lethal dose of 8 Gy gamma radiation. For mice, 30 days after exposure to lethal dose, the survival fraction for the control group was 40% while the 2[nd], 3[rd], and 4[th] groups had survival rates of 20%, 33.30%, and 35.20%, respectively. For rats, 30 days after exposure to the lethal dose, the survival fraction for the control group was 40% while the 2[nd], 3[rd], 4[th] and 5[th] groups had survival rates of 20%, 41.6%, 60.0% and 35.7%, respectively. Results indicate that short term exposure to extremely high levels of natural gamma radiation [up to 196 times higher than the normal background] do not lead to induction of survival adaptive response

[ effect of radioactive lantern mantle powder and bentonite-zeoloite minerals on the volume of blood loss, bleeding and clotting time]

Over the past decade the US army has widely studied new technologies for stopping sever hemorrhages and has introduced an effective Zeolite based hemostatic agent. On the other hand, Mortazavi and his colleagues previously reported the bio-stimulatory effects of the topical application of radioactive lantern mantle powder on wound healing. Their subsequent studies showed significant changes in some histological parameters concerning healing. In this light, here the bio-stimulatory effect of burned radioactive lantern mantles powder as well as two minerals; bentonite and zeolite arepresented. This experimental study was conducted in the center for radiological studies, Shiraz university of Medical Sciences in 2008. Fifty male Wistar rats were divided randomly into 5 groups of 10 animals each. Following anesthesia, animals' tails were cut at a thickness of 5 mm by using a surgical scissor. No intervention was made on the animals of the 1[st] group. The 2[nd] group to 4[th] group received topical non-radioactive lantern mantle powder, radioactive lantern mantle powder, Bentonite mineral or a mixture of Bentonite-Zeoliteat minerals respectively. After treatment with above mentioned agents, the volume of blood loss was measured using a scaled test-tube. The bleeding time and clotting time were also measured using a chronometer. SPSS software was used for statistical analysis. ANOVA was used for comparing the means of each parameter in the 5 groups. The volume of blood loss, bleeding and clotting times in control animals were 4.39 +/- 1.92 cc, 112.10 +/- 39.60 sec and 94.9 +/- 54.26 sec, respectively. In the 5[th] group in which the animals were treated with a mixture of Bentonite-Zeoliteat minerals, the volume of blood loss, bleeding and clotting times were 1.31 +/- 0.60 cc, 34.50 +/- 4.65 sec and 24.2 +/- 4.61 sec, respectively. This is the 1[st] investigation that studied the alterations of bleeding and clotting times following the use of lantern mantle powder as well as bentonite or the mixture of bentonite-zeolite minerals. The results obtained in this study clearly show the significant effect of the volume of blood loss as well as the bleeding time following the topical use of the mixture of bentonite-zeolite minerals. The radioactive and non-radioactive lantern mantle and the bentonite mineral alone had considerable effects but overally the magnitude of the effect was less than that of the mixture of bentonite-zeolite minerals